塞来昔布抑制人类三阴性乳腺癌裸鼠移植瘤生长的实验研究

 【摘要】　目的　探讨塞来昔布（celecoxib）对人类三阴性乳腺癌（TNBC）肿瘤生长及细胞凋亡的影响。方法　32只裸鼠于背部皮下接种人类TNBC细胞株MDA-MB-231，随机分为空白对照组及低、中、高剂量塞来昔布组（25、50、100 mg?kg-1?d-1）。实验结束后，留取移植瘤标本，观察用药前后裸鼠肿瘤体积的变化；流式细胞术（FCM）检测肿瘤细胞凋亡率；免疫组织化学法检测NF-κB p65和p50分子的表达；Western blot法检测凋亡相关分子Caspase-3、Survivin蛋白的表达。结果　塞来昔布治疗组肿瘤体积较对照组均明显减小。中、高剂量塞来昔布治疗组凋亡率分别为（13.58±3.16）％、（21.91±4.75）％，与对照组的（3.15±1.73）％相比差异有统计学意义（t＝6.736，12.151，均P＜0.05），塞来昔布低、中、高剂量组p65表达阳性率分别为79.3 %、46.7 %、23.9 %，与对照组（89.7 %）相比差异有统计学意义（χ2＝3.312，10.785，15.900，均P＜0.05）。Western blot结果显示，塞来昔布治疗后肿瘤组织中Caspase-3蛋白出现了裂解片段，并且随药物浓度增加，裂解片段表达量逐渐增加。Survivin蛋白随药物浓度增加表达逐渐下调。结论　塞来昔布可以诱导TNBC裸鼠移植瘤细胞凋亡，抑制肿瘤生长，其抗肿瘤作用机制可能部分与抑制p65分子以及下调Survivin蛋白表达有关。

Effects of celecoxib on tumor growth and cell apoptosis in human triple-negative breast cancer xenografts in nude mice

Objective To evaluate the effects of celecoxib on tumor growth and cell apoptosis in human triple-negative breast cancer (TNBC) xenografts in nude mice.Methods Human TNBC MDA-MB-231 cells were inoculated subcutaneously into BALB/c nude mice.The mice (n=32) were then randomly divided into 4 groups,the control group and the celecoxib group (receiving 25,50,100 mg·kg-1·d-1 respectively).At the end of the study,tumor tissues were collected and tumor volume was measured.Cell apoptosis was determined by flow cytometry (FCM) analysis.NF-κB p65 and pS0 protein levels were measured by immunohistochemistry.Caspase-3 and survivin protein levels were detected by western blotting.Results celecoxib at dose of 25,50 and 100 mg·kg-1·d-1 inhibited the tumor growth significantly,compared with the control group.FCM results showed that apoptotic rates were (13.58±3.16) ％ and (21.91±4.75) ％ in moderate and high dose of celecoxib-treated group,compared with (3.15±1.73) ％ in control group (t =6.736,P ＜ 0.05;t =12.151,P ＜ 0.05).p65 expressions were 79.3 ％,46.7 ％ and 23.9 ％ in low,moderate and high dose of celecoxib-treated group,compared with 89.7 ％ in control group (x2 =3.312,P ＜ 0.05; x2 =10.785,P ＜ 0.05;x2 =15.900,P ＜ 0.05).Besides,western blotting analysis demonstrated that celecoxib significantly downregulated survivin expression,while upregulated the active form of caspase-3 expression.Conclusion Celecoxib could suppress TNBC tumor growth and induce cell apoptosis,which might be partially associated with inactivation of p65 and downregulation of survivin.